CORESTA Congress, Paris, 2006, SSPOST 08

A cell-free method to assess the gaseous phase cytotoxicity of cigarette smoke

Altadis Research Centre, Fleury-les-Aubrais, France
In order to increase our efficiency in assessing the cigarette smoke's toxicity, we developed fast cell-free methods allowing a first selection before in vitro classical testings. We will here describe a cell-free method, based on the depletion of an anti-oxydant (GSH), applied to an aliquot of PBS-trapped gaseous phase (as required in recommended in vitro testing), that enables to quickly predict the gaseous phase cytotoxicity of cigarette smoke, as classically assessed by Neutral Red test. The correlation between this method and the survival/viability test (Neutral Red cytotoxicity) is excellent (r2 = 0.94). Previously, we had developed a fast method to assess the oxidative effect of the gaseous phase of cigarette smoke(1), by direct bubbling in a GSH solution. We do not have a good correlation between those two free-cell methods (r2 = 0.52). Moreover, the GSH depletions obtained with the "bubbling" method are always higher than the depletions obtained with the "aliquot" method. In order to know the differences between the two cell-free methods, the determination of the products formed by reactive smoke components with GSH in both methods has been also performed. The results clearly show that some oxidant components of the gaseous phase are not trapped in the PBS solution. In conclusion, we will share a new method to quickly predict the gaseous phase cytotoxicity as assessed by Neutral Red test after trapping in PBS solution. We will also demonstrate that several compounds of the native gaseous phase are not tested in this classical configuration.1. Oxydative effect of cigarette smoke: assessment by a fast cell-free method. Xavier Cahours*, Damien Renault, Matthieu Dubois, Véronique Marchand, Bernard Duméry. CORESTA 2004, Kyoto, Japan.